Materials and Methods for Treating Coagulation Disorders

ABSTRACT

This invention is drawn to compounds which are more easily metabolized by the metabolic drug detoxification systems. Particularly, warfarin analogs which have been designed to include esters within the structure of the compounds are taught. The invention teaches methods of reducing the toxicity of drugs comprising the introduction of ester groups into drugs during the synthesis of the drug. This invention is also drawn to methods of treating coagulation disorders comprising the administration of compounds which have been designed to be metabolized by serum or intracellular hydrolases and esterases. Pharmaceutical compositions of the ester containing warfarin, analogs are also taught.

CROSS-REFERENCE TO RELATED APPLICATION(S)

This application is a continuation-in-part of U.S. application Ser. No.10/132,750, filed Apr. 24, 2002; which claims the benefit of U.S.Provisional Application No. 60/286,079, filed Apr. 24, 2001.

BACKGROUND OF INVENTION

Warfarin (coumarin) is an anticoagulant which acts by inhibiting vitaminK-dependent coagulation factors. Warfarin based compounds are,typically, derivatives of 4-hydroxycoumarin, such as3-(a-acetonylbenzyl)-4-hydroxycoumarin (COUMADIN). COUMADIN and othercoumarin anticoagulants inhibit the synthesis of vitamin K dependentclotting factors, which include Factors II, VII, 1× and X. Anticoagulantproteins C and S are also inhibited by warfarin anticoagulants. Warfarinis believed to interfere with clotting factor synthesis by inhibitingvitamin K, epoxide regeneration.

An anticoagulation effect is generally seen about 24 hours afteradministration and single doses of warfarin are effective for 2 to 5days. While anticoagulants have no direct effect on an establishedthrombus and do not reverse ischemic tissue damage, anticoagulanttreatment is intended to prevent the extension of formed clots and/or toprevent secondary thromboembolic complications. These complications mayresult in serious and possibly fatal sequelae.

Warfarin is typically used for the treatment of in patients sufferingfrom atrial fibrillation. Such treatment reduces the incidence ofsystemic thromboembolism and stroke. The FDA has approved warfarin forthe following indications: 1) the treatment or prophylaxis of venousthrombosis and pulmonary embolism, 2) thromboembolic complicationsassociated with atrial fibrillation and/or cardiac valve replacement,and 3) reducing the risk of death, recurring myocardial infarction, andstroke or systemic embolism after myocardial infarction.

A number of adverse effects are associated with the administration ofwarfarin. These include fatal or nonfatal hemorrhage from any tissue ororgan and hemorrhagic complications such as paralysis. Other adverseeffects include paresthesia, headache, chest abdomen, joint, muscle orother pain, dizziness, shortness of breath, difficult breathing orswallowing, unexplained swelling, weakness, hypotension, or unexplainedshock. Other adverse reactions reported includehypersensitivity/allergic reactions, systemic cholesterolmicroembolization, purple toes syndrome, hepatitis, cholestatic hepaticinjury, jaundice, elevated liver enzymes, vasculitis, edema, fever,rash, dermatitis, including bullous eruptions, urticaria, abdominal painincluding cramping, flatulence/bloating, fatigue, lethargy, malaise,asthenia, nausea, vomiting, diarrhea, pain, headache, dizziness, tasteperversion, pruritus, alopecia, cold intolerance, and paresthesiaincluding feeling cold and chills.

Drug toxicity is an important consideration in the treatment of humansand animals. Toxic side effects resulting from the administration ofdrugs include a variety of conditions which range from low grade feverto death. Drug therapy is justified only when the benefits of thetreatment protocol outweigh the potential risks associated with thetreatment. The factors balanced by the practitioner include thequalitative and quantitative impact of the drug to be used as well asthe resulting outcome if the drug is not provided to the individual.Other factors considered include the physical condition of the patient,the disease stage and its history of progression, and any known adverseeffects associated with a drug.

It is important to note that drug toxicity is an important considerationin the treatment of individuals. Toxic side effects resulting from theadministration of drugs include a variety of conditions which range fromlow grade fever to death. Drug therapy is justified only when thebenefits of the treatment protocol outweigh the potential risksassociated with the treatment. The factors balanced by the practitionerinclude the qualitative and quantitative impact of the drug to be usedas well as the resulting outcome if the drug is not provided to theindividual. Other factors considered include the clinical knowledge ofthe patient, the disease and its history of progression, and any knownadverse effects associated with a drug.

Drug elimination is the result of metabolic activity upon the drug andthe subsequent excretion of the drug from the body. Metabolic activitycan take place within the vascular supply and/or within cellularcompartments or organs. The liver is a principal site of drugmetabolism. The metabolic process can be broken down into synthetic andnonsynthetic reactions. In nonsynthetic reactions, the drug ischemically altered by oxidation, reduction, hydrolysis, or anycombination of the aforementioned processes. These processes arecollectively referred to as Phase I reactions.

In Phase II reactions, also known as synthetic reactions orconjugations, the parent drug, or intermediate metabolites thereof, arecombined with endogenous substrates to yield an addition or conjugationproduct. Metabolites formed in synthetic reactions are, typically, morepolar and biologically inactive. As a result, these metabolites are moreeasily excreted via the kidneys (in urine) or the liver (in bile).Synthetic reactions include glucuronidation, amino acid conjugation,acetylation, sulfoconjugation, and methylation.

Drug therapy using warfarin is particularly difficult because themetabolism of warfarin is complex and subject to interactions with ahost of other drugs, including drugs that are commonly prescribed inpatients suffering from atrial fibrillation, such as amiodarone forexample. Warfarin is a mixture of enantiomers having different intrinsicactivities at the vitamin K epoxide reductase (VKER) enzyme. Theseenantiomers have different metabolic pathways using different CYP450isozymes. The CYP450 metabolic system is highly inducible or repressibleby a host of external factors such as diet and other medications. Also,the CYP450 system is subject to many genetic variations and has a lowcapacity and is easily saturable. For these reasons the metabolism ofwarfarin is subject to unpredictable variations and each enantiomer hasa different metabolic fate and different potencies at the VKER enzyme.

In addition, warfarin activity at the VKER enzyme results in inhibitionof coagulation factors II, VII, IX, and X, which have differenthalf-lives of their own, ranging from hours (factor VII) to days (factorX). Because of this complex situation, the pharmacological effect(increased coagulation time) of warfarin becomes apparent only 5 to 10days after a dose. It is therefore easy to understand why warfarintherapy is extremely difficult to predict and why patients are at highrisk of bleeding complications including death. In the current state ofwarfarin therapy, patients on warfarin must report to a coagulation labonce a week in order to be monitored and in order to detect any earlyrisk of bleeding complications. Even with this strict monitoring system,many patients on warfarin die every year from bleeding complications.

The potential clinical problems and business risk associated withdeveloping drugs, which must past through the P450 metabolism“gauntlet”, is markedly increased in the United States by the followingtwo facts: 1) the number of prescriptions filled in this country hasincreased to about 3 billion per year or 10 per person, and 2) patients,particularly those that live longer and have more complex medicalproblems, tend to take multiple medications. The latter issue isimportant because the incidence of ADRs rises exponentially whensubjects take more than four drugs. Although it is good practice toavoid polypharmacy, in many cases this is not possible because patientsrequire different classes of drugs to effectively treat complex medicalconditions.

The landscape of drug R&D is littered by failed drugs that werewithdrawn by the FDA because they caused fatal ADRs involving CYPmetabolism. These drugs were clinically effective and in many casescommercially successful. Notable drugs that were withdrawn due toADR-related deaths involving CYP450 metabolism include terfenadine(February 1998), astemizole (July 1999) and cisapride (January 2000). Ineach of these cases, drug interactions involving CYP3A4 causedconcentrations of the pharmaceutical agent to increase to such a degreethat it significantly inhibited a particular type of potassium channelin the heart called I_(Kr), which in turn, prolonged the QT interval andcaused a potentially fatal form of ventricular tachyarrhythmia calledtorsades de pointes.

A warfarin analog that has a controllable and a predictable metabolicfate, not depending on CYP450, is therefore highly desirable and wouldbe an important addition to the armamentarium of drugs available fortreating atrial fibrillation patients.

BRIEF SUMMARY

The subject invention provides materials and methods for safe andeffective anticoagulant treatment. In a preferred embodiment, thesubject invention provides therapeutic anticoagulant compounds. Thecompounds of the subject invention can be used to treat at-riskpopulations thereby bringing relief of symptoms, improving the qualityof life, preventing acute and long-term complications, reducingmortality and treating accompanying disorders.

Advantageously, the subject invention provides compounds which arereadily metabolized by the physiological metabolic drug detoxificationsystems. Specifically, in a preferred embodiment, the therapeuticcompounds of the subject invention contain an ester group, which doesnot detract from the ability of these compounds to provide a therapeuticbenefit, but which makes these compounds more susceptible to degradationby hydrolases, particularly serum and/or cytosolic esterases. Thesubject invention further provides methods of treatment comprising theadministration of these compounds to individuals in need ofanticoagulant treatment.

In a further embodiment, the subject invention pertains to the breakdownproducts which are formed when the therapeutic compounds of the subjectinvention are acted upon by esterases. These breakdown products can beused as described herein to monitor the clearance of the therapeuticcompounds from a patient.

In yet a further embodiment, the subject invention provides methods forsynthesizing the therapeutic compounds of the subject invention.

The subject invention provides materials and methods for the treatmentof coagulation disorders. Specifically, the subject invention providescompounds which are readily metabolized by the metabolic drugdetoxification systems. Specifically, this invention provides compoundswhich are susceptible to degradation by hydrolases, particularly serumand/or cytosolic esterases. This invention is also drawn to methods oftreating coagulation disorders.

This invention is drawn to compounds which are more easily metabolizedby the metabolic drug detoxification systems. This invention is alsodrawn to methods of treating coagulation disorders. Specifically, thisinvention provides analogs of drugs which have been designed to be moresusceptible to degradation by hydrolases, particularly serum and/orcytosolic esterases and methods of treatment comprising theadministration of these analogs to individuals.

DETAILED DISCLOSURE

The subject invention provides materials and methods for anticoagulanttreatment. Advantageously, the therapeutic compounds of the subjectinvention are stable in storage but have a shorter half-life in thephysiological environment than other drugs which are available foranticoagulant treatment; therefore, the compounds of the subjectinvention can be used with a lower incidence of side effects andtoxicity.

In a preferred embodiment of the subject invention, therapeuticcompounds are provided which are useful in providing anticoagulanttreatment and which contain an ester group that is acted upon byesterases thereby breaking down the compound and facilitating itsefficient removal from the treated individual. In a preferred embodimentthe therapeutic compounds are metabolized by the Phase I drugdetoxification system.

A further aspect of the subject invention pertains to the breakdownproducts that are produced when the therapeutic compounds of the subjectinvention are acted upon by esterases. The presence of these breakdownproducts in the urine or serum can be used to monitor the rate ofclearance of the therapeutic compound from a patient.

The subject invention further provides methods of synthesizing theunique and advantageous therapeutic compounds of the subject invention.Particularly, methods of producing less toxic therapeutic agentscomprising introducing ester groups into therapeutic agents (targetdrugs) are taught. The ester linkage may be introduced into the compoundat a site that is convenient in the manufacturing process for the targetdrug. Additionally, the sensitivity of the ester linkage may bemanipulated by the addition of side groups which hinder or promote thehydrolytic activity of the hydrolases or esterases responsible forcleaving the drug at the ester locus. Methods of adding such sidegroups, as well as the side groups themselves, are well known to theskilled artisan and can be readily carried out utilizing the guidanceprovided herein.

The subject invention further provides anticoagulant treatmentcomprising the administration of a therapeutically effective amount ofesterified coumarin analogs to an individual in need of treatment.Accordingly, the subject invention provides esterified coumarin analogsand pharmaceutical compositions of these esterified compounds. In apreferred embodiment the patient is a human; however, animals also canbe treated.

Adverse drug-drug interactions (DDI), elevation of liver function test(LFT) values, and QT prolongation leading to torsades de pointes (TDP)are three major reasons why drug candidates fail to obtain FDA approval.All these causes are, to some extent metabolism-based. A drug that hastwo metabolic pathways, one oxidative and one non-oxidative, built intoits structure is highly desirable in the pharmaceutical industry. Analternate, non-oxidative metabolic pathway provides the treated subjectwith an alternative drug detoxification pathway (an escape route) whenone of the oxidative metabolic pathways becomes saturated ornon-functional. While a dual metabolic pathway is necessary in order toprovide an escape metabolic route, other features are needed to obtaindrugs that are safe regarding DDI, TDP, and LFT elevations.

In addition to having two metabolic pathways, the drug should have arapid metabolic clearance (short metabolic half-life) so that bloodlevels of unbound drug do not rise to dangerous levels in cases of DDIat the protein level. Also, if the metabolic half-life of the drug istoo long, then the CYP450 system again becomes the main eliminationpathway, thus defeating the original purpose of the design. In order toavoid high peak concentrations and rapidly declining blood levels whenadministered, such a drug should also be administered using a deliverysystem that produces constant and controllable blood levels over time.

In various embodiments, the primary metabolites of the inventivecompounds, regardless of the electrophysiological properties of theparent drug, has, or have, negligible inhibitory activity at the I_(KR)(HERG) channel at normal therapeutic concentrations of the drug inplasma. In other words, the concentration of the metabolite can be atleast five times higher than the normal therapeutic concentration of theparent compound before activity at the I_(KR) channel is observed.Preferably, the concentration of the metabolite is at least ten timeshigher than the normal therapeutic concentration of the parent compoundbefore activity at the I_(KR) channel is observed.

Compounds according to the invention are, primarily, metabolized byendogenous hydrolytic enzymes via hydrolysable bonds engineered intotheir structures. The primary metabolites resulting from this metabolicpathway are water soluble and do not have, or show a reduced incidenceof, DDI when administered with other medications (drugs). Non-limitingexamples of hydrolysable bonds that can be incorporated into compoundsaccording to the invention include amide, ester, carbonate, phosphate,sulfate, urea, urethane, glycoside, or other bonds that can be cleavedby hydrolases.

Additional modifications of the compounds disclosed herein can readilybe made by those skilled in the art. Thus, analogs and salts of theexemplified compounds are within the scope of the subject invention.With a knowledge of the compounds of the subject invention skilledchemists can use known procedures to synthesize these compounds fromavailable substrates. As used in this application, the term “analogs”refers to compounds which are substantially the same as another compoundbut which may have been modified by, for example, adding additional sidegroups. The term “analogs” as used in this application also may refer tocompounds which are substantially the same as another compound but whichhave atomic or molecular substitutions at certain locations in thecompound.

Analogs of the exemplified compounds can be readily prepared usingcommonly known, standard reactions. These standard reactions include,but are not limited to, hydrogenation, methylation, acetylation, andacidification reactions. For example, new salts within the scope of theinvention can be made by adding mineral acids, e.g., HCl H₂SO₄, etc., orstrong organic acids, e.g., formic, oxalic, etc., in appropriate amountsto form the acid addition salt of the parent compound or its derivative.Also, synthesis type reactions may be used pursuant to known proceduresto add or modify various groups in the exemplified compounds to produceother compounds within the scope of the invention.

In a preferred embodiment, the subject invention provides compoundshaving Formula I:

Wherein:

X is independently in each occurrence hydrogen, alkyl, cycloalkyl,halogen, heterocyclyl, hydroxy, alkoxy, R₂, heteroaryl or aryloptionally substituted with COOR₁, or other group, including, forexample, halogens.

R₁ is independently in each occurrence hydrogen, alkyl or allcylaryl,all optionally substituted with lower alkyl, hydroxy, halogen, oralkoxy.

R₂ is

Y is independently in each occurrence (CHR₃)_(n)COOR₄ or aryl optionallysubstituted with COOR₅, wherein n=1 to 3.

R₃ is independently in each occurrence hydrogen, alkyl or alkylaryl,aryl all optionally substituted with lower alkyl, hydroxy, halogen, oralkoxy.

R₄ is independently in each occurrence hydrogen, alkyl or alkylaryl,aryl all optionally substituted with lower allyl, hydroxy, halogen, oralkoxy.

R₅ is independently in each occurrence hydrogen, alkyl or alkylaryl,aryl all optionally substituted with lower alkyl, hydroxy, halogen, oralkoxy.

X and Y taken together can form butyrolactone when X is OH and Y isO-benzoic acid.

Reference herein to “lower alkyl” refers to C₁₋₈ alkyl. As used herein,“aryl” refers to any aromatic group. As set forth herein, the aryl groupmay be substituted or unsubstituted. Possible substituents include, butare not limited to, lower alkyl, hydroxyl, halogen, and alkoxy.

Specifically exemplified herein are the following compounds:

-   3-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-3-phenyl-propionic acid-   3-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-3-phenyl-propionic acid methyl    ester-   3-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-3-phenyl-propionic acid ethyl    ester-   3-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-3-phenyl-propionic acid n-propyl    ester-   3-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-3-phenyl-propionic acid n-butyl    ester-   3-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-3-phenyl-propionic acid 2-butyl    ester-   3-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-3-phenyl-propionic acid    isopropyl ester-   3-Cyclohexyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid-   3-Cyclohexyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid    methyl ester-   3-Cyclohexyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid    ethyl ester-   3-Cyclohexyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid    n-propyl ester-   3-Cyclopentyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid-   3-Cyclopentyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid    methyl ester-   3-Cyclopentyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid    ethyl ester-   3-Cyclopentyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid    n-propyl ester-   3-Propyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid-   3-Propyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid methyl    ester-   3-Propyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid ethyl    ester-   3-Propyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid n-propyl    ester

-   (4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid-   (4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid methyl ester-   (4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid ethyl ester-   (4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid n-propyl ester-   (4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid 2-propyl ester-   (4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid n-butyl ester-   2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid-   2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid methyl ester-   2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid ethyl ester-   2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid n-propyl ester-   2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid isopropyl ester-   2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid n-butyl ester

-   4-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid-   4-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid methyl ester-   4-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid ethyl ester-   4-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid n-propyl    ester-   4-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid n-butyl ester-   2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid-   2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid methyl ester-   2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid ethyl ester-   2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid n-propyl    ester-   2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid n-butyl ester-   3-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid-   3-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid methyl ester-   3-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid ethyl ester-   3-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid n-propyl    ester-   3-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid n-butyl ester

-   2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid-   2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid methyl    ester-   2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid ethyl    ester-   2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid n-propyl    ester-   2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid n-butyl    ester-   2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-butyric acid-   2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-butyric acid methyl ester-   2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-butyric acid ethyl ester-   2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-butyric acid n-propyl    ester-   2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-butyric acid n-butyl ester

-   4-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoic    acid-   4-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoic    acid methyl ester-   4-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoic    acid ethyl ester-   4-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoic    acid n-propyl ester-   [(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoic    acid n-butyl ester-   3-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoic    acid-   3-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoic    acid methyl ester-   3-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoic    acid ethyl ester-   3-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoic    acid n-propyl ester-   3-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoic    acid n-butyl ester-   2-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoic    acid-   2-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoic    acid methyl ester-   2-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoic    acid ethyl ester-   2-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoic    acid n-propyl ester-   2-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoic    acid n-butyl ester

-   (1-Hydroxy-3-oxo-3,4-dihydro-naphthalen-2-yl)-3H-isobenzofuran-1-one

In Formulae II-VII “R” is defined consistent with the exemplifiedcompounds, as well as including the various groups included in thedefinition of “X” in Formula I.

Further specific embodiments of the subject invention include thefollowing compounds:

Advantageously, these halogenated compounds are less favorablesubstrates for cytochrome P450 than their unhalogenated analogs. Theyare therefore more likely to be metabolized by esterases, which isdesirable for eliminating drug-drug interactions according to thesubject invention.

The subject invention also provides processes for the manufacturing ofnovel coumarin derivatives. Examples of synthetic schemes are asfollows:

Scheme 1 provides an exemplary synthesis of C-3 substituted4-hydroxycoumarins. Appropriately substituted bromoacetate and4-hydroxycoumarin in the presence of a base give mixture of O and C₁₋₃alkylated 4-hydroxy coumarins, which are readily separable.

Scheme 2 provides an alternative synthesis of C-3 substituted4-hydroxycoumarins when R1 is aryl groups. 4-hyroxycoumarin and anaromatic aldehyde can be heated in a mixture of triethylamine and formicacid (2:5 molar ratio) to give 3-benzyl-4-hydroxycoumarin, which was inturn treated with 2.2 eq. of BuLi and quenched with carbon dioxide togive coumarin substituted phenyl-acetic acid. Corresponding esters canbe obtained by treating the acid with various alcohols in the presenceof concentrated sulfuric acid.

Scheme 3 illustrates the synthesis of chromen-3-yl-propionic acid.4-hyroxycoumarin, an appropriate aldehyde and meldrum's acid can beheated in EtOH in the presence ammonium acetate to give substitutedpropionate, which can then be treated with 2 eq. of LDA and analkylating agent to provide the chromen-3-yl-propionionates.

Exemplary reaction schemes for the production of derivatives of7-hydroxycoumarin which have anti-coagulant properties are providedbelow. The synthesis of4-Hydroxy-3-(3-methoxy-3-oxo-1-phenylpropyl)-2H-1-benzopyran-2-one 3 isperformed by Michael condensation of 4-Hydroxycoumarin 1 and methyltrans-cinnamate 2 in absolute ethanol in the presence of sodium ethoxideat reflux temperature for 16 hours.

Knoevenagel reaction between 4-Hydroxycoumarin 1 and benzaldehyde 4 inthe presence of piperidinium benzoate gives the benzal adduct 5. Michaeladdition between 5 and ethoxycarbonylmethyldimethylsulfide in toluene inthe presence of DBU as a base gives the cyclopropane derivative 6.Michael addition between 5 and diethyl malonate in absolute ethanol withsodium ethoxide gives 7.

The subject invention further pertains to enantiomerically isolatedcompounds, and compositions comprising the compounds, for the treatmentof coagulation disorders. The isolated enantiomeric forms of thecompounds of the invention are substantially free from one another(i.e., in enantiomeric excess). In other words, the “R” forms of thecompounds are substantially free from the “S” forms of the compounds andare, thus, in enantiomeric excess of the “S” forms. Conversely, “S”forms of the compounds are substantially free of “R” forms of thecompounds and are, thus, in enantiomeric excess of the “R” forms. In oneembodiment of the invention, the isolated enantiomeric compounds are atleast about in 80% enantiomeric excess. In a preferred embodiment, thecompounds are in at least about 90% enantiomeric excess. In a morepreferred embodiment, the compounds are in at least about 95%enantiomeric excess. In an even more preferred embodiment, the compoundsare in at least about 97.5% enantiomeric excess. In a most preferredembodiment, the compounds are in at least 99% enantiomeric excess.

The subject invention also provides methods for treating coagulationdisorders comprising the administration of a therapeutically effectiveamount of the esterified warfarin analogs of this invention to anindividual in need of treatment. The wafarin analogs of this inventionhave applicability in both veterinary and human clinical contexts.Further, the compounds of this invention have therapeutic propertiessimilar to those of the umnodified parent compound (COUMADINE).Accordingly, dosage rates and routes of administration of the disclosedcompounds are similar to those already used in the art and known to theskilled artisan (see, for example, Physicians' Desk Reference, 54^(th)Ed., Medical Economics Company, Montvale, N.J., 2000 or U.S. Pat. No.5,856,525 hereby incorporated by reference in its entirety).

The compounds of the subject invention can be formulated according toknown methods for preparing pharmaceutically useful compositions.Formulations are described in detail in a number of sources which arewell known and readily available to those skilled in the art. Forexample, Remington's Pharmaceutical Science by E. W. Martin describesformulations which can be used in connection with the subject invention.In general, the compositions of the subject invention will be formulatedsuch that an effective amount of the bioactive compound(s) is combinedwith a suitable carrier in order to facilitate effective administrationof the composition.

In accordance with the invention, pharmaceutical compositionscomprising, as an active ingredient, an effective amount of one or moreof the compounds and one or more non-toxic, pharmaceutically acceptablecarrier or diluent. Examples of such carriers for use in the inventioninclude ethanol, dimethyl sulfoxide, glycerol, silica, alumina, starch,and equivalent carriers and diluents.

Further, acceptable carriers can be either solid or liquid. Solid formpreparations include powders, tablets, pills, capsules, cachets,suppositories and dispersible granules. A solid carrier can be one ormore substances which may act as diluents, flavoring agentssolubilizers, lubricants, suspending agents, binders, preservatives,tablet disintegrating agents or an encapsulating material.

The disclosed pharmaceutical compositions may be subdivided into unitdoses containing appropriate quantities of the active component. Theunit dosage form can be a packaged preparation, such as packetedtablets, capsules, and powders in paper or plastic containers or invials or ampoules. Also, the unit dosage can be a liquid basedpreparation or formulated to be incorporated into solid food products,chewing gum, or lozenge.

The term “individual(s)” is defined as a single mammal to which isadministered a compound of the present invention. The mammal may be arodent, for example a mouse or rat, pig, horse, rabbit, goat, pig, cow,cat, dog, or human. In a preferred embodiment, the individual is ahuman.

Following are examples which illustrate procedures for practicing theinvention. These examples should not be construed as limiting. Allpercentages are by weight and all solvent mixture proportions are byvolume unless otherwise noted.

EXAMPLE 1 Preparation of 2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyricacid ethyl ester

To a solution of 4-hydroxy-chromen-2-one (2.0 g) and ethyl2-bromobutyrate (2 ml) in DMF was added anhydrous potassium carbonate(8.5 g). The resulting reaction mixture was stirred at room temperaturefor 72 hours, then diluted with water and extracted with EtOAc. Theorganic layer was washed with saturated aqueous sodium bicarbonate,dried over MgSO₄ and conc. in vacuo to provide colorless oil, which waspurified by column chromatography to give2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid ethyl ester as a whitesolid, MS: 275[M-H].

EXAMPLE 2 Preparation of2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid ethyl ester

A solution of 4-hydroxy-chromen-2-one (2.0 g), aqueous formaldehyde(37%, 0.37 g), Meldrum's acid (1.77 g) and ammonium acetate (0.95 g) inethanol (75 mL) was heated to reflux for 6 hours, then cooled to roomtemperature. The reaction mixture was conc. in vacuo to give the crudeas yellow oil, which was purified by column chromatography to provide3-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid ethyl ester ascolorless oil (1.2 g).

To a solution of 3-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-propionic acidethyl ester in THF at −78° C. was added LDA (1.5M, 3.05 mL) dropwise. Ayellow precipitate was formed during the addition. The reaction wasstirred at −78° C. for 15 min and allowed to warmed to 0° C. and stirredfor 30 min, after which BnBr (0.24 mL in THF) was added dropwise. Thereaction was warmed to room temperature, stirred for 12 hours, cooled to0° C. and quenched with saturated ammonium chloride and extracted withEtOAc. The organic layer was washed with brine, dried over MgSO₄ andconc. in vacuo to a crude colorless oil, which was purified by columnchromatography to provide2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid ethyl esteras a colorless oil (250 mg). MS: 351[M-H].

EXAMPLE 3 Preparation of2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid

2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid ethyl ester(160 mg) in EtOH (1 mL) was added 1N NaOH (1.36 mL). The resultingmixture was heated to 50° C. and stirred for 2 hours, cooled to roomtemperature, acidified with Conc. HCl/ice and extracted with ether. Theorganic layer was dried over MgSO₄ and conc. in vacuo to give2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid as a paleyellow solid (120 mg). MS: 323[M-H].

EXAMPLE 4 Preparation of (4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-aceticacid ethyl ester

Triethylammonium formate (TEAF) was prepared by adding TEA (20.0 mL) toformic acid (16.5 mL) with ice cooling. To TEAF was added benzaldehyde(3.78 mL) and 4-hydroxy-chromen-2-one (6.0 g) and the resulting mixturewas heated to 130-140° C. for 3 hours, cooled to room temperature,diluted with water, and extracted with EtOAc. The organic layer waswashed with brine, dried over MgSO₄ and conc. in vacuo to give a lightyellow solid. The crude solid was recrystallized from EtOH to give3-Benzyl-4-hydroxy-chromen-2-one as a white solid (1.95 g).

To a solution of 3-Benzyl-4-hydroxy-chromen-2-one (2.0 g) in THF at −78°C. was added BuLi (1.6M, 11.4 mL) dropwise during which a yellowprecipitate was formed. The reaction was stirred at −78° C. for 30 minand carbon dioxide gas was bubbled through for 10 min, warmed to 0° C.and quenched with saturated ammonium chloride, extracted with EtOAc(3×50 mL). The aqueous phase was acidified with conc. HCl and extractedwith EtOAc. The organic layer was dried over MgSO₄ and conc. in vacuo toprovide a colorless oil, which crystallize up standing to give(4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid as a white solid(920 mg). MS: 295[M-H].

A solution of (4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid inEtOH with a catalytic amount of conc. sulfuric acid was heated to refluxfor 5 hours, cooled to room temperature, diluted with water andextracted with EtOAc. The organic layer was dried over MgSO₄ and conc.in vacuo to give (4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acidethyl ester as colorless oil, which crystallize upon standing (910 mg).MS: 323[M-H].

EXAMPLE 5 Preparation of4-Hydroxy-3-(3-oxo-1,3-dihydro-isobenzofuran-1-yl)-chromen-2-one

A solution of 4-hydroxy-chromen-2-one (650 mg) and2-carboxybenzyladehyde (300 mg) in EtOH was heated to reflux for 4hours, cooled to room temperature then concentrated in vacuo to give acrude oil, which was diluted with water. The precipitated4-hydroxy-chromen-2-one was collected by filtration (490 mg). A secondcrop of solid was collected from the mother liquor and triturated withhot EtOAc and filtered to provide4-Hydroxy-3-(3-oxo-1,3-dihydro-isobenzofuran-1-yl)-chromen-2-one aswhite solid. MS: 293 [M-H].

EXAMPLE 6 Preparation of2-(4-Hydroxy-2-oxo-2H-chromen-3-Almethyl)-benzoic acid methyl ester

To a solution4-Hydroxy-3-(3-oxo-1,3-dihydro-isobenzofuran-1-yl)-chromen-2-one (60 mg)in ethanol was added 10% Pd/C (10 mg) then stirred under a hydrogenballoon for 12 hours. The reaction mixture was filtered through a pad ofcelite and the filtrate was concentrated in vacuo to give2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid as white solid(50 mg). MS: 295[M-H].

A solution of 2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid inMeOH with a catalytic amount of conc. sulfuric acid was heated to refluxfor 5 hours, cooled to room temperature, diluted with water andextracted with EtOAc. The organic layer was dried over MgSO₄ and conc.in vacuo to give 2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acidmethyl ester as white solid. MS: 309[M-H].

EXAMPLE 7 Mice In Vivo, Bleeding Time

Test substance was administered orally (30, 10 and 3 mg/kg) to a groupof 3 ICR derived male or female mice weighing 22±2 grams, respectively,at 18, 24 and 30 hours before standardized transection of the tip (0.5mm) of each tail. The mice, in holders, were immediately suspendedvertically with the distal 2 cm of each tail immersed in a test tubecontaining saline at 37° C. The time required for beginning a 15 secondperiod of bleeding cessation is then determined; a maximum cut-off timeof 180 seconds is used. Prolongation of bleeding time by 50 percent ormore (≧50%) relative to a control group of animals was consideredsignificant.

EXAMPLE 8 Selected Compounds of the Subject Invention

The subject invention is demonstrated in the production of warfarinanalogs which have been designed to be metabolized by esterase enzymes.Exemplary compounds include structures of the formula:

R₁, R₂ and R₃ can occupy any position on the ring; andare, independently, H, Cl, F, I, lower alkoxy or substituted loweralkoxy, CN, NO₂, NH₂, or —COOR₄where R₄ is lower alkyl or substituted alkyl.Or, either 2 of R₁, R₂, and R₃ are independently methylene, methyne, O,S, NH and together are part of a 5-7-membered cyclic structure, whereinthe cyclic structure can be substituted as defined above with respect toY.Preferred salts are sodium salts.Specific compounds are as follows:

where R₄ is fluorinated or chlorinated lower alkyl having 1-10 carbonatoms (preferred is 1-6 carbon atoms).

Additional compounds are:

wherein

R₁ is selected from the group consisting of —CH₂—COO—R₅, —CH(COOR₅)₂;

R₂ is H;

R₃ is selected from the group consisting of C₁₋₄ alkyl, phenyl, andbenzyl; and,

R₄ is H or a halogen; and,

R₅ is selected from the group consisting of C₁₋₄ alkyl, phenyl, andbenzyl groups.

Other embodiments of this invention contemplate compounds of theformulae:

wherein R is selected from the group consisting of C₁₋₄ alkyl, phenyl,and benzyl groups and R₄ is defined as above, H or a halogen.

Further specific embodiments include the following:

EXAMPLE 9 Halogenated Compounds

In certain embodiments, the subject invention provides compounds havingFormula I:

wherein:

R₁ is H or CH₂COOH;

R₂, R₃, and R₄ are independently H, Cl, OCF₃, or COOR₅, in which atleast one of R₂, R₃, and R₄ is COOR₅, where R₅ is a halogenated alkyl,cycloalkyl, heteroalkyl, heterocycloalkyl, aryl, or heteroaryl group,comprising at least 1 halogen atom into its structure, preferablyfluorine or chlorine, and which can be optionally substituted with otherhalogens atoms, OH, O-alkyl, or O-fluorinated alkyl.Alternatively, R₂ and R₃ together can form an aromatic structure to giveFormula II:

wherein:

R₁ is H or CH₂COOH; R₄ is H, Cl, OCF₃; and

R₆, R₇, and R₈ are independently H, Cl, OCF₃, or COOR₉, in which atleast one of R₆, R₇, and R₈ is COOR₉, where R₉ is a halogenated alkyl,cycloalkyl, heteroalkyl, heterocycloalkyl, aryl, or heteroaryl group,comprising at least 1 halogen atom into its structure, preferablyfluorine or chlorine, and which can be optionally substituted with otherhalogens atoms, OH, O-alkyl, or O-fluorinated alkyl.

The subject invention also provides compounds of Formula I and FormulaII wherein R₅ and R₉ are H. Compounds where R₅ and R₉ are H are theprimary metabolites when compounds of Formula I and II are administeredto a mammal, including human. They are essentially devoid of activity atthe VKER enzyme, but they are useful for monitoring drug levels inpatients.

Specific embodiments of the present invention include the followingcompounds:

Advantageously, these halogenated compounds are less favorablesubstrates for cytochrome CYP450 than their unhalogenated analogs. Theyare therefore more likely to be metabolized by esterases, which isdesirable for eliminating drug-drug interactions according to thesubject invention.

The subject invention also provides processes for the manufacturing ofthe novel compounds. The synthesis of these compounds can be achieved asshown in schemes 1 and 2.

In scheme 1, 4-hydroxycoumarin and a substituted aromatic aldehyde canbe heated in a mixture of triethylamine and formic acid (2:5 molarratio) to give the correspondingly substituted3-benzyl-4-hydroxycoumarin wherein R₁ is hydrogen. Scheme 2 describesthe synthetic pathway where R₁ is CH₂COOH. In scheme 2,4-hydroxycoumarin, an appropriately substituted aromatic aldehyde, andmeldrum's acid can be heated in ethanol in the presence of ammoniumacetate to give the correspondingly substituted chromen-3-yl-propionate,which in turn can be hydrolyzed using a base such as NaOH followed byacidification in order to provide the chromen-3-yl-propionic acid whereR₂, R₃, and R₄ are defined as above.

The subject invention further pertains to enantiomerically purecompounds, and compositions comprising the compounds, for the treatmentof coagulation disorders. The isolated enantiomeric forms of thecompounds of the invention are substantially free from one another(i.e., in enantiomeric excess). In other words, the “R” forms of thecompounds are substantially free from the “S” forms of the compounds andare, thus, in enantiomeric excess of the “S” forms. Conversely, “S”forms of the compounds are substantially free of “R” forms of thecompounds and are, thus, in enantiomeric excess of the “R” forms. In oneembodiment of the invention, the isolated enantiomeric compounds are atleast about in 80% enantiomeric excess. In a preferred embodiment, thecompounds are in at least about 90% enantiomeric excess. In a morepreferred embodiment, the compounds are in at least about 95%enantiomeric excess. In an even more preferred embodiment, the compoundsare in at least about 97.5% enantiomeric excess. In a most preferredembodiment, the compounds are in at least 99% enantiomeric excess.

Modifications of the compounds disclosed herein can readily be made bythose skilled in the art. Thus, analogs, derivatives, and salts of theexemplified compounds are within the scope of the subject invention.With a knowledge of the compounds of the subject invention, and theirstructures, skilled chemists can use known procedures to synthesizethese compounds from available substrates.

All patents, patent applications, provisional applications, andpublications referred to or cited herein are incorporated by referencein their entirety, including all figures and tables, to the extent theyare not inconsistent with the explicit teachings of this specification.Any compounds specifically disclosed in Synthetic Communications Journal(1993) 25:631-640 are specifically excluded from the scope of thecompounds of the subject invention.

It should be understood that the examples and embodiments describedherein are for illustrative purposes only and that various modificationsor changes in light thereof will be suggested to persons skilled in theart and are to be included within the spirit and purview of thisapplication.

1.-3. (canceled)
 4. A method for providing anticoagulant activity to apatient in need of such treatment, the method comprising administering acompound or pharmaceutically acceptable salt thereof, where the compoundis3-(1-Hydroxy-3-oxo-3,4-dihydro-naphthalen-2-yl)-3H-isobenzofuran-1-one;3-Cyclohexyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid;3-Cyclohexyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid methylester; 3-Cyclohexyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acidethyl ester; 3-Cyclohexyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionicacid n-propyl ester;3-Cyclopentyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid;3-Cyclopentyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid methylester; 3-Cyclopentyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acidethyl ester; 3-Cyclopentyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionicacid n-propyl ester;3-Propyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid;3-Propyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid methylester; 3-Propyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid ethylester; 3-Propyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acidn-propyl ester; (4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid;(4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid methyl ester;(4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid ethyl ester;(4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid n-propyl ester;(4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid 2-propyl ester;(4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid n-butyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid;2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid methyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid ethyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid n-propyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid isopropyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid n-butyl ester;4-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid;4-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid methyl ester;4-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid ethyl ester;4-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid n-propyl ester;4-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid n-butyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid;2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid methyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid ethyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid n-propyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid n-butyl ester;3-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid;3-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid methyl ester;3-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid ethyl ester;3-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid n-propyl ester;3-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid n-butyl ester;2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid;2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid methylester; 2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid ethylester; 2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acidn-propyl ester; 2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionicacid n-butyl ester; 2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-butyricacid; 2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-butyric acid methylester; 2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-butyric acid ethylester; 2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-butyric acid n-propylester; 2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-butyric acid n-butylester;4-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid methyl ester;4-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid ethyl ester;4-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid n-propyl ester;4-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid n-butyl ester;3-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid;3-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid methyl ester;3-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid ethyl ester;3-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid n-propyl ester;3-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid n-butyl ester;2-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid;2-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid methyl ester;2-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid ethyl ester;2-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid n-propyl ester; or2-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid n-butyl ester.
 5. A method according to claim 4, wherein thecompound is3-(1-Hydroxy-3-oxo-3,4-dihydro-naphthalen-2-yl)-3H-isobenzofuran-1-one,or pharmaceutically acceptable salts thereof.
 6. A method according toclaim 4, wherein the compound is3-Cyclohexyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid;3-Cyclohexyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid methylester; 3-Cyclohexyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acidethyl ester; 3-Cyclohexyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionicacid n-propyl ester;3-Cyclopentyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid; orpharmaceutically acceptable salts thereof.
 7. A method according toclaim 4, wherein the compound is3-Cyclopentyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid methylester; 3-Cyclopentyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acidethyl ester; 3-Cyclopentyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionicacid n-propyl ester;3-Propyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid;3-Propyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid methylester; or pharmaceutically acceptable salts thereof.
 8. A methodaccording to claim 4, wherein the compound is3-Propyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid ethyl ester;3-Propyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid n-propylester; (4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid;(4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid methyl ester;(4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid ethyl ester; orpharmaceutically acceptable salts thereof.
 9. A method according toclaim 4, wherein the compound is(4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid n-propyl ester;(4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid 2-propyl ester;(4-Hydroxy-2-oxo-2H-chromen-3-yl)-phenyl-acetic acid n-butyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid;2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid methyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid ethyl ester; orpharmaceutically acceptable salts thereof.
 10. A method according toclaim 4, wherein the compound is2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid n-propyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid isopropyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid n-butyl ester;4-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid;4-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid methyl ester;4-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid ethyl ester; orpharmaceutically acceptable salts thereof.
 11. A method according toclaim 4, wherein the compound is2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid n-propyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid isopropyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-yl)-butyric acid n-butyl ester;4-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid;4-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid methyl ester;4-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid ethyl ester; orpharmaceutically acceptable salts thereof.
 12. A method according toclaim 4, wherein the compound is4-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid n-propyl ester;4-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid n-butyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid;2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid methyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid ethyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid n-propyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid n-butyl ester;3-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid;3-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid methyl ester;3-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid ethyl ester; orpharmaceutically acceptable salts thereof.
 13. A method according toclaim 4, wherein the compound is3-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid n-propyl ester;3-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-benzoic acid n-butyl ester;2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid;2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid methylester; 2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acid ethylester; 2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionic acidn-propyl ester; 2-Benzyl-3-(4-hydroxy-2-oxo-2H-chromen-3-yl)-propionicacid n-butyl ester; 2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-butyricacid; 2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-butyric acid methylester; or pharmaceutically acceptable salts thereof.
 14. A methodaccording to claim 4, wherein the compound is2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-butyric acid ethyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-butyric acid n-propyl ester;2-(4-Hydroxy-2-oxo-2H-chromen-3-ylmethyl)-butyric acid n-butyl ester;4-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid methyl ester;4-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid ethyl ester;4-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid n-propyl ester;4-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid n-butyl ester;3-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid; or pharmaceutically acceptable salts thereof.
 15. A methodaccording to claim 4, wherein the compound is3-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid methyl ester;3-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid ethyl ester;3-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid n-propyl ester;3-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid n-butyl ester;2-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid;2-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid methyl ester;2-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid ethyl ester;2-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid n-propyl ester;2-[(4-Hydroxy-2-oxo-2H-chromen-3-yl)-(4-hydroxy-1-oxo-1H-isochromen-3-yl)-methyl]-benzoicacid n-butyl ester; or pharmaceutically acceptable salts thereof.